Locomotion of free-swimming ghost knifefish: anal fin kinematics during four behaviors.
The maneuverability demonstrated by the weakly electric ghost knifefish (Apteronotus albifrons) is a result of its highly flexible ribbon-like anal fin, which extends nearly three-quarters the length of its body and is composed of approximately 150 individual fin rays. To understand how movement of the anal fin controls locomotion we examined kinematics of the whole fin, as well as selected individual fin rays, during four locomotor behaviors executed by free-swimming ghost knifefish: forward swimming, backward swimming, heave (vertical) motion, and hovering. We used high-speed video (1000 fps) to examine the motion of the entire anal fin and we measured the three-dimensional curvature of four adjacent fin rays in the middle of the fin during each behavior to determine how individual fin rays bend along their length during swimming. Canonical discriminant analysis separated all four behaviors on anal fin kinematic variables and showed that forward and backward swimming behaviors contrasted the most: forward behaviors exhibited a large anterior wavelength and posterior amplitude while during backward locomotion the anal fin exhibited both a large posterior wavelength and anterior amplitude. Heave and hover behaviors were defined by similar kinematic variables; however, for each variable, the mean values for heave motions were generally greater than for hovering. Individual fin rays in the middle of the anal fin curved substantially along their length during swimming, and the magnitude of this curvature was nearly twice the previously measured maximum curvature for ray-finned fish fin rays during locomotion. Fin rays were often curved into the direction of motion, indicating active control of fin ray curvature, and not just passive bending in response to fluid loading.


Estimating population parameters using the structured serial coalescent with Bayesian MCMC inference when some demes are hidden.
Using the structured serial coalescent with Bayesian MCMC and serial samples, we estimate population size when some demes are not sampled or are hidden, ie ghost demes. It is found that even with the presence of a ghost deme, accurate inference was possible if the parameters are estimated with the true model. However with an incorrect model, estimates were biased and can be positively misleading. We extend these results to the case where there are sequences from the ghost at the last time sample. This case can arise in HIV patients, when some tissue samples and viral sequences only become available after death. When some sequences from the ghost deme are available at the last sampling time, estimation bias is reduced and accurate estimation of parameters associated with the ghost deme is possible despite sampling bias. Migration rates for this case are also shown to be good estimates when migration values are low.


Surgical Results and Outcomes After Reimplantation for the Management of Anomalous Aortic Origin of the Right Coronary Artery.
Anomalous aortic origin of the right coronary artery (AAORCA) has been reported to cause myocardial ischemia, leading to angina, dyspnea, and decreased exercise tolerance. Reimplantation is a repair technique devised to exclude the abnormal intramural portion of the anomalous artery and avoid the known late attrition of saphenous vein grafts. Our study aims to evaluate the medium-term clinical outcomes with this technique. A retrospective review was made of patients who underwent repair of AAORCA by reimplantation between 2002 and 2014 in two institutions in Western Australia. Follow-up computed tomography coronary angiography was used to assess the status of the reimplanted right coronary artery (RCA). Data on survival, freedom from symptoms, cardiac events, and cardiac interventions were also analyzed. Of the 16 patients (aged 17 to 70 years old), 14 (88%) were symptomatic before surgery, with angina (50%) and exertional dyspnea (56%) being the most common symptoms. Surgical reimplantation was successful in 15 patients (94%) without operative mortality. One patient required saphenous vein bypass grafting of the RCA intraoperatively after presumed failed repair and difficulty weaning from cardiopulmonary bypass. All patients who had successful reimplantation of AAORCA were symptom-free after surgery, and none had subsequent cardiac events attributable to the RCA or required further interventions. Ten patients (67%) had computed tomography coronary angiography after surgery; none had stenosis, kinking, or compression of the RCA by the pulmonary artery. Two further patients (including the patient who underwent saphenous vein grafting for presumed failed reimplantation) underwent conventional angiography, which demonstrated patent reimplantations. To the best of our knowledge, this is the largest reported series of anomalous RCA managed by surgical reimplantation. Our results suggest that this technique is safe and has excellent medium to long-term results regarding symptom-free survival.


Development of a predictive pharmacokinetic model for a novel cyclooxygenase-2 inhibitor.
A predictive population pharmacokinetic model was developed for a novel cyclooxygenase-2 (COX-2) inhibitor CS-706, using data from 130 subjects in 3 phase 1 trials after single or multiple doses of CS-706 (2- to 800-mg doses daily, up to 14 days) and validated using sparse data from a separate study. A 2-compartment model described the data. Typical apparent clearance (CL/F) was 47.2 L/h and was reduced by 43% at doses greater than 200 mg. Apparent clearance was decreased by 38% in female subjects and by 64% and 15%, respectively, in poor/intermediate CYP 2D6 and poor CYP 2C9 metabolizers. Typical apparent volume of the central compartment was 166 L and increased with body weight. Bioavailability increased by 42% after nighttime doses and decreased saturably with increasing dose (50% reduction at 221 mg). Predicted exposures in Japanese subjects were reduced relative to whites because of a lower frequency of poor metabolizers. The model may aid in optimizing the design of future studies and predicting exposures in other subpopulations.


Information in the sample covariate distribution in prevalent cohorts.
Methods of estimation and inference about survival distributions based on length-biased samples are well-established. Comparatively little attention has been given to the assessment of covariate effects in the context of length-biased samples, but prevalent cohort studies often have this objective. We show that, like the survival distribution, the covariate distribution from a prevalent cohort study is length-biased, and that this distribution may contain parametric information about covariate effects on the survival time. As a result, a likelihood based on the joint distribution of the survival time and the covariates yields estimates of covariate effects which are at least as efficient as estimates arising from a traditional likelihood which conditions on covariate values in the length-biased sample. We also investigate the empirical bias of estimators arising from a joint likelihood when the population covariate distribution is misspecified. The asymptotic relative efficiencies and empirical biases under model misspecification are assessed for both proportional hazards and accelerated failure time models. The various methods considered are applied in an illustrative analysis of risk factors for death following onset of dementia using data collected in the Canadian Study of Health and Aging.


Multiple Hodgkin lymphoma-associated loci within the HLA region at chromosome 6p21.3.
Since an association between the human leukocyte antigen (HLA) region and Hodgkin lymphoma (HL) was first reported in 1967, many studies have reported associations between HL risk and both single nucleotide polymorphism (SNP) and classic HLA allele variation in the major histocompatibility complex. However, population stratification and the extent and complexity of linkage disequilibrium within the major histocompatibility complex have hindered efforts to fine-map causal signals. Using SNP data to impute alleles at classic HLA loci, we have conducted an integrated analysis of HL risk within the HLA region in 582 early-onset HL cases and 4736 controls. We confirm that the strongest signal of association comes from an SNP located in the class II region, rs6903608 (odds ratio [OR] = 1.79, P = 6.63 × 10(-19)), which is unlikely to be driven by association to HLA-DRB, DQA, or DQB alleles. In addition, we identify independent signals at rs2281389 (OR = 1.73, P = 6.31 × 10(-13)), a SNP that maps closely to HLA-DPB1, and the class II HLA allele DQA1*02:01 (OR = 0.56, P = 1.51 × 10(-7)). These data suggest that multiple independent loci within the HLA class II region contribute to the risk of developing early-onset HL.


Differential response of type C and intracisternal type A particle markers in cells treated with iododeoxyuridine and dexamethasone.
Mouse neuroblastoma cells containing intracisternal type A particles were treated with iododeoxyuridine and dexamethasone to induce the release of type C oncornavirus particles. For 5 days after treatment, antigenic markers and DNA polymerase activities specific to particles of each of the two types were assayed in the cells and in pellets obtained by high-speed centrifugation of the culture fluid. There was a marked release of C-particle antigen (p30) and DNA polymerase activity in extracellular particulate form, reaching a maximum on day 3 after treatment and falling thereafter. In contrast, no extracellular A-particle antigen was detected, and A-particle-specific DNA polymerase activity in the medium pellets did not increase from the original very low level. Electron microscopy confirmed the presence of free type C virus particles, but not intracisternal type A particles, in the culture fluid. Although intracellular levels of C-particle antigen rose 20- to 30-fold per milligram of cell protein, intracellular A-particle antigen and DNA polymerase activity did not vary more than two-fold. The relative rate of A-particle synthesis in the treated cells, as judged by incorporation of radioactive amino acids into the major structural protein (P73), was also unchanged over the period of observation. Thus, the induction of type C virus particle formation in cultured neuroblastoma cells had no detectable effect on the quantity, synthesis rate, or location of intracisternal type A particles.


The importance of lung cancer screening with low-dose computed tomography for Medicare beneficiaries.
The National Lung Screening Trial has provided convincing evidence of a substantial mortality benefit of lung cancer screening with low-dose computed tomography (CT) for current and former smokers at high risk. The United States Preventive Services Task Force has recommended screening, triggering coverage of low-dose CT by private health insurers under provisions of the Affordable Care Act. The Centers for Medicare & Medicaid Services (CMS) are currently evaluating coverage of lung cancer screening for Medicare beneficiaries. Since 70% of lung cancer occurs in patients 65 years or older, CMS should cover low-dose CT, thus avoiding the situation of at-risk patients being screened up to age 64 through private insurers and then abruptly ceasing screening at exactly the ages when their risk for developing lung cancer is increasing. Legitimate concerns include false-positive findings that lead to further testing and invasive procedures, overdiagnosis (detection of clinically unimportant cancers), the morbidity and mortality of surgery, and the overall costs of follow-up tests and procedures. These concerns can be mitigated by clear criteria for screening high-risk patients, disciplined management of abnormalities based on algorithms, and high-quality multidisciplinary care. Lung cancer screening with low-dose CT can lead to early diagnosis and cure for thousands of patients each year. Professional societies can help CMS responsibly implement a program that is patient-centered and minimizes unintended harms and costs.


Attention and interhemispheric transfer: a behavioral and fMRI study.
When both detections and responses to visual stimuli are performed within one and the same hemisphere, manual reaction times (RTs) are faster than when the two operations are carried out in different hemispheres. A widely accepted explanation for this difference is that it reflects the time lost in callosal transmission. Interhemispheric transfer time can be estimated by subtracting RTs for uncrossed from RTs for crossed responses (crossed-uncrossed difference, or CUD). In the present study, we wanted to ascertain the role of spatial attention in affecting the CUD and to chart the brain areas whose activity is related to these attentional effects on interhemispheric transfer. To accomplish this, we varied the proportion of crossed and uncrossed trials in different blocks. With this paradigm subjects are likely to focus attention either on the hemifield contralateral to the responding hand (blocks with 80% crossed trials) or on the ipsilateral hemifield (blocks with 80% uncrossed trials). We found an inverse correlation between the proportion of crossed trials in a block and the CUD and this effect can be attributed to spatial attention. As to the imaging results, we found that in the crossed minus uncrossed subtraction, an operation that highlights the neural processes underlying interhemispheric transfer, there was an activation of the genu of the corpus callosum as well as of a series of cortical areas. In a further commonality analysis, we assessed those areas which were activated specifically during focusing of attention onto one hemifield either contra- or ipsilateral to the responding hand. We found an activation of a number of cortical and subcortical areas, notably, parietal area BA 7 and the superior colliculi. We believe that the main thrust of the present study is to have teased apart areas important in interhemispheric transmission from those involved in spatial attention.


Long noncoding RNA in prostate, bladder, and kidney cancer.
Genomic regions without protein-coding potential give rise to millions of protein-noncoding RNA transcripts (noncoding RNA) that participate in virtually all cellular processes. Research over the last 10 yr has accumulated evidence that long noncoding RNAs (lncRNAs) are often altered in human urologic cancers. To review current progress in the biology and implication of lncRNAs associated with prostate, bladder, and kidney cancer. The PubMed database was searched for articles in the English language with combinations of the Medical Subject Headings terms long non coding RNA, long noncoding RNA, long untranslated RNA, cancer, neoplasms, prostate, bladder, and kidney. We summarise existing knowledge on the systematics, biology, and function of lncRNAs, particularly these involved in prostate, kidney, and bladder cancer. We also discuss the possible utilisation of lncRNAs as novel biomarkers and potential therapeutic targets in urologic malignancies and portray the major challenges and future perspectives of ongoing lncRNA research. LncRNAs are important regulators of gene expression interacting with the major pathways of cell growth, proliferation, differentiation, and survival. Alterations in the function of lncRNAs promote tumour formation, progression, and metastasis of prostate, bladder, and kidney cancer. LncRNAs can be used as noninvasive tumour markers in urologic malignancies. Increased knowledge of the molecular mechanisms by which lncRNAs perform their function in the normal and malignant cell will lead to a better understanding of tumour biology and could provide novel therapeutic targets for the treatment of urologic cancers. In this paper we reviewed current knowledge of long noncoding RNAs (lncRNAs) for the detection and treatment of urologic cancers. We conclude that lncRNAs can be used as novel biomarkers in prostate, kidney, or bladder cancer. LncRNAs hold promise as future therapeutic targets, but more research is needed to gain a better understanding of their biologic function.


Taste at first (person) sight: Visual perspective modulates brain activity implicitly associated with viewing unhealthy but not healthy foods.
Every day, people are exposed to images of appetizing foods that can lead to high-calorie intake and contribute to overweight and obesity. Research has documented that manipulating the visual perspective from which eating is viewed helps resist temptation by altering the appraisal of unhealthy foods. However, the neural basis of this effect has not yet been examined using neuroimaging methods. Moreover, it is not known whether the benefits of this strategy can be observed when people, especially overweight, are not explicitly asked to imagine themselves eating. Last, it remains to be investigated if visual perspective could be used to promote healthy foods. The present work manipulated camera angles and tested whether visual perspective modulates activity in brain regions associated with taste and reward processing while participants watch videos featuring a hand grasping (unhealthy or healthy) foods from a plate during functional magnetic resonance imagining (fMRI). The plate was filmed from the perspective of the participant (first-person perspective; 1PP), or from a frontal view as if watching someone else eating (third-person perspective; 3PP). Our findings reveal that merely viewing unhealthy food cues from a 1PP (vs. 3PP) increases activity in brain regions that underlie representations of rewarding (appetitive) experiences (amygdala) and food intake (superior parietal gyrus). Additionally, our results show that ventral striatal activity is positively correlated with body mass index (BMI) during exposure to unhealthy foods from a 1PP (vs. 3PP). These findings suggest that unhealthy foods should be promoted through third-person (video) images to weaken the reward associated with their simulated consumption, especially amongst overweight people. It appears however that, as such, manipulating visual perspective fails to enhance the perception of healthy foods. Their promotion thus requires complementary solutions.


SNPs associated with cerebrospinal fluid phospho-tau levels influence rate of decline in Alzheimer's disease.
Alzheimer's Disease (AD) is a complex and multifactorial disease. While large genome-wide association studies have had some success in identifying novel genetic risk factors for AD, case-control studies are less likely to uncover genetic factors that influence progression of disease. An alternative approach to identifying genetic risk for AD is the use of quantitative traits or endophenotypes. The use of endophenotypes has proven to be an effective strategy, implicating genetic risk factors in several diseases, including anemia, osteoporosis and heart disease. In this study we identify a genetic factor associated with the rate of decline in AD patients and present a methodology for identification of other such factors. We have used an established biomarker for AD, cerebrospinal fluid (CSF) tau phosphorylated at threonine 181 (ptau(181)) levels as an endophenotype for AD, identifying a SNP, rs1868402, in the gene encoding the regulatory sub-unit of protein phosphatase B, associated with CSF ptau(181) levels in two independent CSF series (P(combined) = 1.17 x 10(-05)). We show no association of rs1868402 with risk for AD or age at onset, but detected a very significant association with rate of progression of disease that is consistent in two independent series (P(combined) = 1.17 x 10(-05)). Our analyses suggest that genetic variants associated with CSF ptau(181) levels may have a greater impact on rate of progression, while genetic variants such as APOE4, that are associated with CSF Aβ(42) levels influence risk and onset but not the rate of progression. Our results also suggest that drugs that inhibit or decrease tau phosphorylation may slow cognitive decline in individuals with very mild dementia or delay the appearance of memory problems in elderly individuals with low CSF Aβ(42) levels. Finally, we believe genome-wide association studies of CSF tau/ptau(181) levels should identify novel genetic variants which will likely influence rate of progression of AD.


Pleiotropic Effects of Myocardial MMP-9 Inhibition to Prevent Ventricular Arrhythmia.
Observational studies have established a strong association between matrix metalloproteinase-9 (MMP-9) and ventricular arrhythmia. However, whether MMP-9 has a causal link to ventricular arrhythmia, as well as the underlying mechanism, remains unclear. Here, we investigated the mechanistic involvement of myocardial MMP-9 in the pathophysiology of ventricular arrhythmia. Increased levels of myocardial MMP-9 are linked to ventricular arrhythmia attacks after angiotensin II (Ang II) treatment. MMP-9-deficient mice were protected from ventricular arrhythmia. Increased expressions of protein kinase A (PKA) and ryanodine receptor phosphorylation at serine 2808 (pS2808) were correlated with inducible ventricular arrhythmia. MMP-9 deficiency consistently prevented PKA and pS2808 increases after Ang II treatment and reduced ventricular arrhythmia. Calcium dynamics were examined via confocal imaging in isolated murine cardiomyocytes. MMP-9 inhibition prevents calcium leakage from the sarcoplasmic reticulum and reduces arrhythmia-like irregular calcium transients via protein kinase A and ryanodine receptor phosphorylation. Human induced pluripotent stem cell-derived cardiomyocytes similarly show that MMP-9 inhibition prevents abnormal calcium leakage. Myocardial MMP-9 inhibition prevents ventricular arrhythmia through pleiotropic effects, including the modulation of calcium homeostasis and reduced calcium leakage.


Prospective Memory in a Language-Trained Chimpanzee (Pan troglodytes).
Prospective memory involves the encoding, retention, and implementation of an intended future action. Although humans show many forms of prospective memory, less is known about the future oriented processes of nonhuman animals, or their ability to use prospective memory. In this experiment, a chimpanzee named Panzee, who had learned to associate geometric forms called lexigrams with real-world referents, was given a prospective memory test. Panzee selected between two foods the one she wanted to receive more immediately. That food was scattered in an outdoor yard where she could forage for it. Also outdoors were lexigram tokens, one of which represented the food item that remained indoors throughout a 30 minute period, and that could be obtained if Panzee brought in the token that matched that food item. After foraging for the selected food item, Panzee consistently remembered to retrieve and return the correct token when food was available indoors, whereas on control trials involving no indoor food she rarely returned a token. This indicated that Panzee encoded information relevant to the future action of token retrieval after extended delays for one type of food, even when a more immediately preferred food was available.


Electron attachment to molecules in a cluster environment.
Low-energy dissociative electron attachment (DEA) to the CF(2)Cl(2) and CF(3)Cl molecules in a water cluster environment is investigated theoretically. Calculations are performed for the water trimer and water hexamer. It is shown that the DEA cross section is strongly enhanced when the attaching molecule is embedded in a water cluster, and that this cross section grows as the number of water molecules in the cluster increases. This growth is explained by a trapping effect that is due to multiple scattering by water molecules while the electron is trapped in the cluster environment. The trapping increases the resonance lifetime and the negative ion survival probability. This confirms qualitatively existing experiments on electron attachment to the CF(2)Cl(2) molecule placed on the surface of H(2)O ice. The DEA cross sections are shown to be very sensitive to the position of the attaching molecule within the cluster and the orientation of the electron beam relative to the cluster.


Generation of an immunodominant CTL epitope is affected by proteasome subunit composition and stability of the antigenic protein.
Generation of the HLA-A0201 (A2) influenza Matrix 58-66 epitope contained within the full-length Matrix protein is impaired in cells lacking the proteasome subunits low molecular protein 2 (LMP2) and LMP7. This Ag presentation block can be relieved by transfecting the wild-type LMP7 cDNA into LMP7-deficient cells. A mutated form of LMP7, lacking the two threonines at the catalytic active site, was equally capable of relieving the block in presentation of the influenza Matrix A2 epitope. These observations were extended by analyzing whether modification of the influenza Matrix protein could overcome the block in presentation of the A2 Matrix epitope. Expression of either a rapidly degraded form of the full-length Matrix protein or shorter Matrix fragments led to an efficient presentation of the A2 influenza Matrix epitope by LMP7-negative cells. These findings demonstrate two main points: 1) LMP7 incorporation into the proteasome is of greater importance for the generation of the influenza A2 Matrix epitope than the presence of the LMP7's catalytic site; and 2) the interplay between cytosolic proteases and stability of target proteins is of importance in optimization of Ag presentation. These observations may have relevance to the immunodominance of tumor and viral epitopes and raise the possibility that generation of shorter protein fragments could be a mechanism to ensure optimal Ag presentation by cells expressing low levels of LMP7.


Antigenic basis of diversity in the gammadelta T cell co-receptor WC1 family.
WC1 co-receptors are transmembrane glycoproteins with 11 extracellular scavenger receptor cysteine rich (SRCR) domains. They are related to the CD163 family but are uniquely expressed by gammadelta T cells. We recently showed that at least 13 members comprise the WC1 gene family in cattle, a model animal species for studies of gammadelta T cell biology. Since WC1 co-receptors participate in directing functional responses by gammadelta T cells either through the ligands they bind or the signals they transduce, availability of reagents to identify the expression of individual WC1 molecules of this diverse family would be valuable in further elucidating mechanisms of gammadelta T cell responsiveness. Although monoclonal antibodies (mAbs) have been widely used to identify WC1 co-receptors on gammadelta T cells, the locations of the antigenic epitopes recognized are unknown. Here, we mapped the epitopes to particular SRCR domains and evaluated their distribution among WC1 molecules. To do this, cDNA representing the extracellular domains of seven different WC1 genes was expressed in mammalian cells and analyzed for reactivity with anti-WC1 mAbs using ELISA and Western blotting. The study included mAbs that are broadly reactive with WC1(+) gammadelta T cells and those that divide WC1(+) gammadelta T cells into functionally distinct subpopulations. We found that mAb CC15 is a pan-reactive anti-WC1 mAb recognizing an epitope in the closely related SRCR domains 2 and 7 and that this epitope is present in at least domain 2 or 7 of all seven WC1 molecules evaluated here. Five other anti-WC1 mAbs, typified by mAb IL-A29, were found to be broadly reactive, recognizing epitopes in the related SRCR domains 4 and 9 but each having a unique pattern of reactivity with the seven WC1 molecules. Finally, the subpopulation-specific anti-WC1 mAbs, including those that recognize either the archetypal WC1.1 or WC1.2 molecule, were found to react with epitopes in the most variable WC1 domain, i.e. domain 1, of a restricted number of WC1 co-receptors.


Emotion Unchained: Facial Expression Modulates Gaze Cueing under Cognitive Load.
Direction of eye gaze cues spatial attention, and typically this cueing effect is not modulated by the expression of a face unless top-down processes are explicitly or implicitly involved. To investigate the role of cognitive control on gaze cueing by emotional faces, participants performed a gaze cueing task with happy, angry, or neutral faces under high (i.e., counting backward by 7) or low cognitive load (i.e., counting forward by 2). Results show that high cognitive load enhances gaze cueing effects for angry facial expressions. In addition, cognitive load reduces gaze cueing for neutral faces, whereas happy facial expressions and gaze affected object preferences regardless of load. This evidence clearly indicates a differential role of cognitive control in processing gaze direction and facial expression, suggesting that under typical conditions, when we shift attention based on social cues from another person, cognitive control processes are used to reduce interference from emotional information.


Earthworms increase plant production: a meta-analysis.
To meet the challenge of feeding a growing world population with minimal environmental impact, we need comprehensive and quantitative knowledge of ecological factors affecting crop production. Earthworms are among the most important soil dwelling invertebrates. Their activity affects both biotic and abiotic soil properties, in turn affecting plant growth. Yet, studies on the effect of earthworm presence on crop yields have not been quantitatively synthesized. Here we show, using meta-analysis, that on average earthworm presence in agroecosystems leads to a 25% increase in crop yield and a 23% increase in aboveground biomass. The magnitude of these effects depends on presence of crop residue, earthworm density and type and rate of fertilization. The positive effects of earthworms become larger when more residue is returned to the soil, but disappear when soil nitrogen availability is high. This suggests that earthworms stimulate plant growth predominantly through releasing nitrogen locked away in residue and soil organic matter. Our results therefore imply that earthworms are of crucial importance to decrease the yield gap of farmers who can't -or won't- use nitrogen fertilizer.


Cigarette smoke primes the pulmonary environment to IL-1α/CXCR-2-dependent nontypeable Haemophilus influenzae-exacerbated neutrophilia in mice.
Cigarette smoke has a broad impact on the mucosal environment with the ability to alter host defense mechanisms. Within the context of a bacterial infection, this altered host response is often accompanied by exacerbated cellular inflammation, characterized by increased neutrophilia. The current study investigated the mechanisms of neutrophil recruitment in a murine model of cigarette smoke exposure and, subsequently, a model of both cigarette smoke exposure and bacterial infection. We investigated the role of IL-1 signaling in neutrophil recruitment and found that cigarette smoke-induced neutrophilia was dependent on IL-1α produced by alveolar macrophages. In addition to being the crucial source of IL-1α, alveolar macrophages isolated from smoke-exposed mice were primed for excessive IL-1α production in response to bacterial ligands. To test the relevance of exaggerated IL-1α production in neutrophil recruitment, a model of cigarette smoke exposure and nontypeable Haemophilus influenzae infection was developed. Mice exposed to cigarette smoke elaborated an exacerbated CXCR2-dependent neutrophilia in response to nontypeable Haemophilus influenzae. Exacerbated neutrophilia was dependent on IL-1α priming of the pulmonary environment by cigarette smoke as exaggerated neutrophilia was dependent on IL-1 signaling. These data characterize a novel mechanism of cigarette smoke priming the lung mucosa toward greater IL-1-driven neutrophilic responses to bacteria, with a central role for the alveolar macrophage in this process.


Behind the Rainbow, "Tongqi" Wives of Men Who Have Sex With Men in China: A Systematic Review.
Due to the restrictions and stigmatization of homosexuality in China, there has emerged the "Tongqi," or the wives of men who have sex with men (MSM). There are around 14 million Tongqi wives whose needs for support are often overshadowed. This phenomenon has been largely under researched, this review is the first to address the current data on the Tongqi. The aim of this systematic review is to begin to provide insight into the pre-existing data and the further support that is needed for the wives of MSM. The researchers searched PubMed, Web of Science, EMBASE, PsycINFO, CNKI, Sinomed and WangFang databases from their inception date until June 7, 2019. Handsearching was also completed to provide a rich data set. The articles were summarized and analyzed for thematic clusters. From the selected article, five themes emerged, including Sexual Health Issues, Intimate Partner Violence, Mental Health Status, Marriage Dissatisfaction, and Coping Strategies. These themes often intersected to provide a complex understanding of the current gaps in support provided to Tongqi. Tongqi wives remain a hidden population in Chinese mainstream society, who deserves a sensitive approach to support. The study revealed that the MSM wives suffer severe mental, physical, health, and life related harms. However, instead of situating them into the victim roles, many women take on an identity of empowerment and are working together, aiming to make social changes. In order to address the Tongqi phenomenon, it is also essential to reduce the discrimination toward homosexuality. Tongqi are a special group of Chinese women, they require further intensive research attention.


Preoperative factors associated with adverse events during awake craniotomy: analysis of 609 consecutive cases.
Awake surgery is becoming more standard and widely practiced for neurosurgical cases, including but not limited to brain tumors. The optimal selection of patients who can tolerate awake surgery remains a challenge. The authors performed an updated cohort study, with particular attention to preoperative clinical and imaging characteristics that may have an impact on the viability of awake craniotomy in individual patients. The authors conducted a single-institution cohort study of 609 awake craniotomies performed in 562 patients. All craniotomies were performed by the same surgeon at Toronto Western Hospital during the period from 2006 to 2018. Analyses of preoperative clinical and imaging characteristics that may have an impact on the viability of awake craniotomy in individual patients were performed. Twenty-one patients were recorded as having experienced intraoperative adverse events necessitating deeper sedation, which made the surgery no longer "awake." In 2 of these patients, conversion to general anesthesia was performed. The adverse events included emotional intolerance of awake surgery (n = 13), air embolism (n = 3), generalized seizure (n = 4), and unexpected subarachnoid hemorrhage (n = 1). Preoperative cognitive decline, dysphasia, and low performance status, as indicated by the Karnofsky Performance Status (KPS) score, were significantly associated with emotional intolerance on univariate analysis. Only a preoperative KPS score < 70 was significantly associated with this event on multivariate analysis (p = 0.0057). Compared with patients who did not experience intraoperative adverse events, patients who did were more likely to undergo inpatient admission (p = 0.0004 for all cases; p = 0.0036 for cases originally planned as day surgery), longer hospital stay (p < 0.0001), and discharge to a location other than home (p = 0.032). Preoperative physical status was found to be the most decisive factor in predicting whether patients can tolerate an awake craniotomy without complications, whereas older age and history of psychiatric treatment were not necessarily associated with adverse events. Patients who had intraoperative adverse events often had reduced chances of same-day discharge and discharge to home. Preoperative careful selection of patients who are most likely to tolerate the procedure is the key to success for awake surgery.


Functional and structural abnormalities of the kidney and urinary tract in severely malnourished children - A hospital based study.
The association of malnutrition and systemic diseases like chronic kidney disease (CKD) is well known. Various urinary tract abnormalities may be associated with malnutrition. So objective of current study was to determine the frequency of functional and structural urinary tract abnormalities in severely malnourished children admitted in Nutritional Rehabilitation Unit (NRU) of a tertiary care facility, Karachi. This descriptive cases series of 78 children was conducted in NRU from October 2014 - March 2015. All newly admitted children aged 2-60 months, diagnosed as Severe Acute Malnutrition (SAM) were studied and children with known kidney and urinary tract disorders were excluded. Detailed history, examination and investigations like serum creatinine, ultrasound kidney and urinary tract in addition to routine tests for SAM, were done. A proforma was used to collect demographic data, clinical history, physical findings, and radio-imaging and biochemical investigations. Glomerular filtration rate (GFR) was calculated using Schwartz equation. Data was analyzed using descriptive statistics. Among 78 children, male to female ratio was equal. Mean age was 18±15.53 months and majority (79.48%) of children were below 24 months. Majority (82%) of children with SAM had marasmus whereas 18% had edematous malnutrition. Out of 78, 57 (73%) children had either functional (80.7%) and or structural (19.3%) abnormalities whereas 21(36.84%) had normal functional and structural status. Most common functional abnormality was subnormal GFR (<90ml/min/1.73 m2) found in all 46 children. Functional abnormities were more common in children below 24 months. Other functional disorders were Bartter syndrome, renal tubular acidosis and urinary tract infection (UTI) found in two cases each. Common structural abnormalities were echogenic kidneys (n=4, 36%), hydronephrosis (n=3, 27%), hypoplastic kidneys (n=3, 27%) and calculi (n=1, 9%). Subnormal GFR was also found in all cases with structural abnormalities. UTI was observed exclusively in two children among 11 with structural abnormalities. A high frequency of functional abnormalities and noticeable proportion of structural abnormalities of urinary tract were detected in children with SAM. Current finding suggest that multicenter study at national level may be undertaken to generate better data about prevalence of renal diseases in SAM.


Back pain and degenerative abnormalities in the spine of young elite divers: a 5-year follow-up magnetic resonance imaging study.
Several studies have been published on disc degeneration among young athletes in sports with great demands on the back, but few on competitive divers; however, there are no long-term follow-up studies. Twenty elite divers between 10 and 21 years of age, with the highest possible national ranking, were selected at random without knowledge of previous or present back injuries or symptoms for an MRI study of the thoraco-lumbar spine in a 5-year longitudinal study. The occurrence of MRI abnormalities and their correlation with back pain were evaluated. Eighty-nine percent of the divers had a history of back pain and the median age at the first episode of back pain was 15 years. Sixty-five percent of the divers had MRI abnormalities in the thoraco-lumbar spine already at baseline. Only one diver without abnormalities at baseline had developed abnormalities at follow-up. Deterioration of any type of abnormality was found in 9 of 17 (53%) divers. Including all disc levels in all divers, the total number of abnormalities increased by 29% at follow-up, as compared to baseline. The most common abnormalities were reduced disc signal, Schmorl's nodes, and disc height reduction. Since almost all divers had previous or present back pain, a differentiated analysis of the relationship between pain and MRI findings was not possible. However, the high frequency of both back pain and MRI changes suggests a causal relationship. In conclusion, elite divers had high frequency of back pain at young ages and they run a high risk of developing degenerative abnormalities of the thoraco-lumbar spine, probably due to injuries to the spine during the growth spurt.


The Precise Breakpoint Mapping in Paracentric Inversion 10q22.2q23.3 by Comprehensive Cytogenomic Analysis, Multicolor Banding, and Single-Copy Chromosome Sequencing.
Detection and precise genomic mapping of balanced chromosomal abnormalities in patients with impaired fertility or a clinical phenotype represent a challenge for current cytogenomics owing to difficulties with precise breakpoint localization in the regions enriched for DNA repeats and high genomic variation in such regions. Here, we present a comprehensive cytogenomic approach to breakpoint mapping in a rare paracentric inversion on 10q (in a patient with oligoasthenoteratozoospermia and necrozoospermia) that does not affect other phenotype traits. Multicolor banding, chromosomal microarray analysis, chromosome microdissection with reverse painting, and single-copy sequencing of the rearranged chromosome were performed to determine the length and position of the inverted region as well as to rule out a genetic imbalance at the breakpoints. As a result, a paracentric 19.251 Mbp inversion at 10q22.2q23.3 was described. The most probable location of the breakpoints was predicted using the hg38 assembly. The problems of genetic counseling associated with enrichment for repeats and high DNA variability of usual breakpoint regions were discussed. Possible approaches for cytogenomic assessment of couples with balanced chromosome rearrangements and problems like reproductive failures were considered and suggested as useful part of effective genetic counseling.


Diversity of physical properties of bacterial extracellular membrane vesicles revealed through atomic force microscopy phase imaging.
Bacteria release nanometer-scale extracellular membrane vesicles (MVs) to mediate a variety of biological processes. We analyzed individual MVs under physiological conditions by phase imaging of high-speed atomic force microscopy to assess the physiological heterogeneity of MVs isolated from bacterial cultures. Phase imaging makes it possible to map the physical properties of an individual, fragile MV in an isolated MV population containing a broad variety of vesicle diameters, from 20 to 150 nm. We also developed a method for quantitatively comparing the physical properties of MVs among samples. This allowed for the comparison of the physical properties of MVs isolated from different bacterial species. We compared bacterial MVs isolated from four bacterial species and artificially synthesized liposomes. We demonstrate that each bacterial species generates physically heterogeneous types of MVs, unlike the physical homogeneity displayed by liposomes. These results indicate that the physical heterogeneity of bacterial MVs is mainly caused by compositional differences mediated through biological phenomena and could be unique to each species. We provide a new methodology using phase imaging that would pave the way for single-vesicle analysis of extracellular vesicles of a broad size range.


The R78K and D117E active-site variants of Saccharomyces cerevisiae soluble inorganic pyrophosphatase: structural studies and mechanistic implications.
We have solved the structure of two active-site variants of soluble inorganic pyrophosphatases (PPase), R78K and D117K, at resolutions of 1.85 and 2.15 A and R-factors of 19.5% and 18.3%, respectively. In the R78K variant structure, the high-affinity phosphate group (P1) is missing, consistent with the wild-type structure showing a bidentate interaction between P1 and Arg78, and solution data showing a decrease in P1 affinity in the variant. The structure explains why the mutation affects P1 and pyrophosphate binding much more than would be expected by the loss of one hydrogen bond: Lys78 forms an ion-pair with Asp71, precluding an interaction with P1. The R78K variant also provides the first direct evidence that the low-affinity phosphate group (P2) can adopt the structure that we believe is the immediate product of hydrolysis, with one of the P2 oxygen atoms co-ordinated to both activating metal ions (M1 and M2). If so, the water molecule (Wat1) between M1 and M2 in wild-type PPase is, indeed, the attacking nucleophile. The D117E variant structure likewise supports our model of catalysis, as the Glu117 variant carboxylate group is positioned where Wat1 is in the wild-type: the potent Wat1 nucleophile is replaced by a carboxylate co-ordinated to two metal ions. Alternative confirmations of Glu117 may allow Wat1 to be present but at much reduced occupancy, explaining why the pKa of the nucleophile increases by three pH units, even though there is relatively little distortion of the active site. These new structures, together with parallel functional studies measuring catalytic efficiency and ligand (metal ion, PPi and Pi) binding, provide strong evidence against a proposed mechanism in which Wat1 is considered unimportant for hydrolysis. They thus support the notion that PPase shares mechanistic similarity with the "two-metal ion" mechanism of polymerases.


Safe use of MRI in people with cardiac implantable electronic devices.
MR scanning in patients with cardiac implantable electronic devices (CIEDs) was formerly felt to be contraindicated, but an increasing number of patients have an implanted MR conditional device, allowing them to safely undergo MR scanning, provided the manufacturer's guidance is adhered to. In addition, some patients with non-MR conditional devices may undergo MR scanning if no other imaging modality is deemed suitable and there is a clear clinical indication for scanning which outweighs the potential risk. The following guidance has been formulated by the British Heart Rhythm Society and endorsed by the British Cardiovascular Society and others. It describes protocols that should be followed for patients with CIEDs undergoing MR scanning. The recommendations, principles and conclusions are supported by the Royal College of Radiologists.


Near Real-Time Surveillance to Assess the Safety of the 9-Valent Human Papillomavirus Vaccine.
Human papillomavirus is the most common sexually transmitted infection in the United States and causes certain anogenital and oropharyngeal cancers. The 9-valent human papillomavirus vaccine (9vHPV) provides protection against additional types not included in the quadrivalent vaccine. We conducted near real-time vaccine safety surveillance for 24 months after the vaccine became available in the Vaccine Safety Datalink. Immunizations and adverse events were extracted weekly from October 2015 to October 2017 from standardized data files for persons 9 to 26 years old at 6 Vaccine Safety Datalink sites. Prespecified adverse events included anaphylaxis, allergic reaction, appendicitis, Guillain-Barré syndrome, chronic inflammatory demyelinating polyneuropathy, injection site reaction, pancreatitis, seizure, stroke, syncope, and venous thromboembolism. The observed and expected numbers of events after 9vHPV were compared weekly by using sequential methods. Both historical and concurrent comparison groups were used to identify statistical signals for adverse events. Unexpected signals were investigated by medical record review and/or additional analyses. During 105 weeks of surveillance, 838 991 doses of 9vHPV were administered. We identified unexpected statistical signals for 4 adverse events: appendicitis among boys 9 to 17 years old after dose 3; pancreatitis among men 18 to 26 years old; and allergic reactions among girls 9 to 17 years old and women 18 to 26 years old after dose 2. On further evaluation, which included medical record review, temporal scan analysis, and additional epidemiological analyses, we did not confirm signals for any adverse events. After 2 years of near real-time surveillance of 9vHPV and several prespecified adverse events, no new safety concerns were identified.


Reliability of Trial Information Across Registries for Trials With Multiple Registrations: A Systematic Review.
Clinical trial registries are important for gaining an overview of ongoing research efforts and for deterring and identifying publication bias and selective outcome reporting. The reliability of the information in trial registries is uncertain. To assess the reliability of information across registries for trials with multiple registrations. For this systematic review, 360 protocols of randomized clinical trials (RCTs) approved by research ethics committees in Switzerland, the UK, Canada, and Germany in 2012 were evaluated. Clinical trial registries were searched from March to September 2019 for corresponding registrations of these RCTs. For RCTS that were recorded in more than 1 clinical trial registry, key trial characteristics that should be identical among all trial registries (ie, sponsor, funding source, primary outcome, target sample size, trial status, date of first patient enrollment, results available, and main publication indexed) were extracted in duplicate. Agreement between the different trial registries for these key characteristics was analyzed descriptively. Data analyses were conducted from May 1 to November 30, 2020. Representatives from clinical trial registries were interviewed to discuss the study findings between February 1 and March 31, 2021. The analysis included 197 RCTs registered in more than 1 trial registry (151 in 2 registries and 46 in 3 registries), with 188 trials in ClinicalTrials.gov, 185 in the European Union Drug Regulating Authorities Clinical Trials Database (EudraCT), 20 in ISRCTN, and 47 in other registries. The agreement of key information across all registries was as follows: 178 of 197 RCTs (90%; 95% CI, 85%-94%) for sponsor, 18 of 20 (90%; 95% CI, 68%-99%) for funding source (funding was not reported on ClinicalTrials.gov), 154 of 197 (78%; 95% CI, 72%-84%) for primary outcome, 90 of 197 (46%; 95% CI, 39%-53%) for trial status, 122 of 194 (63%; 95% CI, 56%-70%) for target sample size, and 43 of 57 (75%; 95% CI, 62%-86%) for the date of first patient enrollment when the comparison time was increased to 30 days (date of first patient enrollment was not reported on EudraCT). For results availability in trial registries, agreement was 122 of 197 RCTs (62%; 95% CI, 55%-69%) for summary data reported in the registry and 91 of 197 (46%; 95% CI, 39%-53%) for whether a published article with the main results was indexed. Different legal requirements were stated as the main reason for inconsistencies by representatives of clinical trial registries. In this systematic review, for a substantial proportion of registered RCTs, information about key trial characteristics was inconsistent across trial registries, raising concerns about the reliability of the information provided in these registries. Further harmonization across clinical trial registries may be necessary to increase their usefulness.


Effect of miR-23 on oxidant-induced injury in human retinal pigment epithelial cells.
Micro(mi)RNAs negatively regulate a wide variety of genes through degradation or posttranslational inhibition of their target genes. The purpose of this study was to investigate the role of miR-23a in modulating RPE cell survival and gene expression in response to oxidative damage. The expression level of miR-23a was measured in macular retinal pigment epithelial (RPE) cells of donor eyes with aged-related macular degeneration (AMD) and age-matched normal eyes by using qRT-PCR. Cultured human ARPE-19 cells were transfected with miR-23a mimic or inhibitor. Cell viability was assessed by the MTT assay. Apoptosis was determined by incubating cells with hydrogen peroxide (H(2)O(2)) or t-butylhydroperoxide (tBH). Caspase-3 activity and DNA fragmentation were measured by enzyme-linked immunosorbent assays. The protein relevant to apoptosis, such as Fas expression level, was analyzed by Western blot analysis. miR-23a expression was significantly downregulated in macular RPE cells from AMD eyes. H(2)O(2)-induced ARPE-19 cell death and apoptosis were increased by an miR-23a inhibitor and decreased by an miR-23a mimic. Computational analysis found a putative target site of miR-23a in the 3'UTR of Fas mRNA, which was verified by a luciferase reporter assay. Forced overexpression of miR-23a decreased H(2)O(2) or tBH-induced Fas upregulation, and this effect was blocked by downregulation of miR-23a. The protection of RPE cells against oxidative damage is afforded by miR-23a through regulation of Fas, which may be a novel therapeutic target in retinal degenerative diseases.


Fast spoiled gradient-recalled MR imaging of thoracic aortic dissection: preliminary clinical experience at 1.5 T.
The purpose of this study was to evaluate fast spoiled gradient-recalled (FSPGR) magnetic resonance (MR) imaging in the diagnosis of thoracic aortic dissection (TAD). Twenty-eight patients with suspected TAD underwent MR imaging with FSPGR and either cine or cardiac-gated spin-echo MR techniques. The average scanning time for the FSPGR images was approximately 1 min. Three readers interpreted the FSPGR images for the presence or absence of TAD. An ROC analysis was done. At a specificity of 90%, the sensitivity ranged from 52% to 90% for the three readers. Pulsatility artifacts and mural thrombus were causes of false-positive and false-negative readings. The areas under the ROC curves (Az) ranged from 0.85 to 0.97 for the three readers. There was a statistically significant difference in the Az values for two of the experienced readers (p = .02). The correct type of dissection was determined in only 65% of the true-positive diagnoses. FSPGR has a very limited role in screening and for rapid evaluation of the unstable patient. The results are reader dependent and susceptible to pulsatility artifacts. Determination of the type of dissection is limited. With a suspected thoracic aortic dissection, therefore, additional imaging sequences should be obtained to maximize accuracy.


A descriptive study of BRCA1 testing and reactions to disclosure of test results.
The identification of the BRCA1 gene is a powerful tool for predicting a patient's lifetime risk for carcinoma of the breast and ovary when she has hereditary breast/ovarian carcinoma (HBOC) syndrome. The process of BRCA1 testing and genetic counseling and participants' reactions to test results, are described. Education about the natural history of HBOC syndrome and the pros and cons of genetic testing was provided to 14 HBOC families comprised of 2549 bloodline relatives. Of these, 388 underwent DNA testing. After informed consent was given by participants, formal linkage analysis and gene mutation studies were performed on the families. Qualitative data on intentions and emotional reactions were collected by physicians/counselors during the genetic counseling sessions. Of those tested, 181 received their results after further genetic counseling. Seventy-eight of them were positive and 100 were negative for BRCA1 gene mutation. Three had ambiguous findings. The most common reasons given for seeking DNA testing were concern about risk to children and concern about surveillance and prevention. Prophylactic mastectomy was considered by 35% of women who tested positive, whereas prophylactic oophorectomy was considered an important option by 76%. Twenty-five percent of both BRCA1 positive and negative individuals were concerned about discrimination by insurance companies. Eighty percent of those who tested negative reported emotional relief, whereas over one-third of those who tested positive reported sadness, anger, or guilt. DNA testing of patients with HBOC syndrome must be performed in the context of genetic counseling. The authors' results demonstrate the many complex clinical and nonclinical issues that are important in this process.


Effects of mobile phone radiation on X-ray-induced tumorigenesis in mice.
The increased use of mobile phones has raised the question of possible health effects of such devices, particularly the risk of cancer. It seems unlikely that the low-level radiofrequency (RF) radiation emitted by them would damage DNA directly, but its ability to act as a tumor promoter is less well characterized. In the current study, we evaluated the effect of low-level RF radiation on the development of cancer initiated in mice by ionizing radiation. Two hundred female CBA/S mice were randomized into four equal groups at the age of 3 to 5 weeks. The mice in all groups except the cage-control group were exposed to ionizing radiation at the beginning of the study and then to RF radiation for 1.5 h per day, 5 days a week for 78 weeks. One group was exposed to continuous NMT (Nordic Mobile Telephones)-type frequency-modulated RF radiation at a frequency of 902.5 MHz and a nominal average specific absorption rate (SAR) of 1.5 W/kg. Another group was exposed to pulsed GSM (Global System for Mobile)-type RF radiation (carrier-wave frequency 902.4 MHz, pulse frequency 217 Hz) at a nominal average SAR of 0.35 W/kg. The control animals were sham-exposed. Body weight, clinical signs, and food and water consumption were recorded regularly. Hematological examinations and histopathological analyses of all lesions and major tissues were performed on all animals. The RF-radiation exposures did not increase the incidence of any neoplastic lesion significantly. We conclude that the results do not provide evidence for cancer promotion by RF radiation emitted by mobile phones.


Hematology of free-ranging, lactating northern fur seals, Callorhinus ursinus.
Thirteen standard hematology values were determined for a healthy and growing population of free-ranging, lactating northern fur seals (Callorhinus ursinus) from Lovushki Island in the Kuril Islands of far-east Russia. Results are presented from 24 females sampled between June and August during the 3-yr period of 2006-08. Hematologic values have been made available for future comparisons with the declining population of northern fur seals on the Pribilof Islands, Alaska, and are compared with published values for other otariid species.


Acute effects of meal fatty acids on postprandial NEFA, glucose and apo E response: implications for insulin sensitivity and lipoprotein regulation?
Our aim was to determine whether meal fatty acids influence insulin and glucose responses to mixed meals and whether these effects can be explained by variations in postprandial NEFA and Apo, which regulate the metabolism of triacylglycerol-rich lipoproteins (Apo C and E). A single-blind crossover study examined the effects of single meals enriched in saturated fatty acids SFA), n-6 PUFA and MUFA on plasma metabolite and insulin responses. The triacylglycerol response following the PUFA meal showed a lower net incremental area under the curve than following the SFA and MUFA meals (P<0.007). Compared with the SFA meal, the PUFA meal showed a lower net incremental area under the curve for the NEFA response from initial suppression to the end of the postprandial period (180-480 min; P<0.02), and both PUFA and MUFA showed a lower net incremental glucose response (P<0.02), although insulin concentrations were similar between meals. The pattern of the Apo E response was also different following the SFA meal (P<0.02). There was a significant association between the net incremental NEFA (180-480 min) and glucose response (rs=0.409, P=0.025), and in multiple regression analysis the NEFA response accounted for 24 % of the variation in glucose response. Meal SFA have adverse effects on the postprandial glucose response that may be due to greater elevations in NEFA arising from differences in the metabolism of SFA- v. PUFA- and MUFA-rich lipoproteins. Elevated Apo E responses to high-SFA meals may have important implications for the hepatic metabolism of triacylglycerol-rich lipoproteins.


Modified Lagrangian method for modeling water quality in distribution systems.
Previous work has shown that Lagrangian methods are more efficient for modeling the transport of chemicals in a water distribution system. Two such methods, the Lagrangian Time-Driven Method (TDM) and Event-Driven Method (EDM) are compared for varying concentration tolerance and computational water quality time step. A new hybrid method (EDMNET) is developed which improves the accuracy of the Lagrangian methods. All the above methods are incorporated in an existing hydraulic simulation model. The integrated model is run for different network problems under varying conditions. The TDM-generated solutions are affected by both concentration tolerance and water quality time step, whereas EDM solutions are dependent on concentration tolerance. The EDMNET solutions are less sensitive to variations in these parameters. The threshold solutions are determined for all the methods and compared. The hybrid method simulates the nodal concentrations accurately with least maximum segmentation of network and reasonable computational effort as compared to the other Lagrangian methods.


Resurrection and redescription of Varestrongylus alces (Nematoda: Protostrongylidae), a lungworm of the Eurasian moose (Alces alces), with report on associated pathology.
Varestrongylus alces, a lungworm in Eurasian moose from Europe has been considered a junior synonym of Varestrongylus capreoli, in European roe deer, due to a poorly detailed morphological description and the absence of a type-series. Specimens used in the redescription were collected from lesions in the lungs of Eurasian moose, from Vestby, Norway. Specimens were described based on comparative morphology and integrated approaches. Molecular identification was based on PCR, cloning and sequencing of the ITS-2 region of the nuclear ribosomal DNA. Phylogenetic analysis compared V. alces ITS-2 sequences to these of other Varestrongylus species and other protostrongylids. Varestrongylus alces is resurrected for protostrongylid nematodes of Eurasian moose from Europe. Varestrongylus alces causes firm nodular lesions that are clearly differentiated from the adjacent lung tissue. Histologically, lesions are restricted to the parenchyma with adult, egg and larval parasites surrounded by multinucleated giant cells, macrophages, eosinophilic granulocytes, lymphocytes. The species is valid and distinct from others referred to Varestrongylus, and should be separated from V. capreoli. Morphologically, V. alces can be distinguished from other species by characters in the males that include a distally bifurcated gubernaculum, arched denticulate crura, spicules that are equal in length and relatively short, and a dorsal ray that is elongate and bifurcated. Females have a well-developed provagina, and are very similar to those of V. capreoli. Morphometrics of first-stage larvae largely overlap with those of other Varestrongylus. Sequences of the ITS-2 region strongly support mutual independence of V. alces, V. cf. capreoli, and the yet undescribed species of Varestrongylus from North American ungulates. These three taxa form a well-supported crown-clade as the putative sister of V. alpenae. The association of V. alces and Alces or its ancestors is discussed in light of host and parasite phylogeny and host historical biogeography. Varestrongylus alces is a valid species, and should be considered distinct from V. capreoli. Phylogenetic relationships among Varestrongylus spp. from Eurasia and North America are complex and consistent with faunal assembly involving recurrent events of geographic expansion, host switching and subsequent speciation.


Prognosis of Asymptomatic Intracranial Stenosis in Patients With Transient Ischemic Attack and Minor Stroke.
Asymptomatic intracranial stenosis (ICS) is a frequent finding on imaging results, particularly in the assessment of acute stroke. Although the management of symptomatic ICS is informed by randomized trials, to our knowledge there are few data on the prevalence and prognosis of asymptomatic ICS in patients with stroke and transient ischemic attack (TIA). To study the age-specific prevalence and prognosis of asymptomatic ICS in a population-based cohort of patients with TIA and minor stroke. All patients (predominantly white) recruited to the Oxford Vascular Study (Oxfordshire, England) between March 1, 2011, and March 1, 2018, with TIA and minor ischemic stroke (National Institutes of Health Stroke Scale score, ≤3), irrespective of age, were included (n = 1579). We determined the age-specific prevalence of 50% or more asymptomatic ICS and the associated stroke risk by face-to-face follow-up to 2018 on standard medical treatment without stenting. Patients underwent magnetic resonance angiography of the intracranial and cervicocranial arteries, computed tomography angiography if magnetic resonance angiography was contraindicated, or carotid/transcranial Doppler ultrasonography if computed tomography angiography was contraindicated. The primary outcomes were the prevalence and prognosis of asymptomatic ICS. Of 1368 eligible patients (mean [SD] age, 69.2 [13.9] years; 700 men [51.2%]) with intracranial vascular imaging, 426 ICS were identified in 260 patients (19.0%): 58 (4.2%) with only symptomatic ICS, 155 (11.3%) with only asymptomatic ICS, and 47 (3.4%) with both. The prevalence of any asymptomatic ICS increased from 4.8% for patients younger than 70 years to 34.6% for patients 90 years or older (P for trend < .001; odds ratio per decade, 1.96; 95% CI, 1.69-2.27) and was greater than that of 50% or more asymptomatic carotid bifurcation stenosis (202 [14.8%] vs 105 patients [7.2%]; relative risk, 2.04; 95% CI, 1.63-2.55, P < .001). However, the 155 patients with only asymptomatic ICS had no increase in risk of ischemic stroke compared with those with no ICS (unadjusted HR, 1.03, 95% CI, 0.49-2.17), with 8 first recurrent events (5.2%) during 506 patient-years of follow-up and 3 in the territory of the ICS (annualized risk, 0.59%; 95% CI, 0.12-1.73). The prevalence of asymptomatic ICS increases with age in white patients with TIA and minor stroke and is greater than that of asymptomatic carotid stenosis, but asymptomatic ICS does not increase the short- or medium-term risk of distal recurrent ischemic stroke for patients receiving standard medical treatment.


The Outcomes Of Serial Cerebrospinal Fluid Removal In Elderly Patients With Idiopathic Normal Pressure Hydrocephalus.
Elderly patients with iNPH (idiopathic normal pressure hydrocephalus) might be potentially high-risk surgical patients. Our purpose was to investigate the outcome of serial removal of cerebrospinal fluid (CSF) in the patients with iNPH who refused to have the ventriculoperitoneal or lumboperitoneal shunt surgery or had contraindications to them. There were 42 patients, with a median age of 78 years. Recurrent CSF removal was performed when the patients had deteriorated gait which was defined as >10% pre-removal change on the average of two walking trials during timed up and go (TUG). All the patients underwent mini-mental status examination (MMSE), frontal assessment battery (FAB), Stroop test, Tinetti Performance Oriented Mobility Assessment (POMA), TUG and nine-hole peg test (NHPT) with the dominant hand, before and after CSF removal. Thirty-five patients had two CSF removal procedures with a mean interim period of 7.4 months ranging from 1.5 to 35 months. Thirteen patients had three CSF removal procedures. The mean TUG scores were decreased after the first, second and third procedures (p<0.001; p<0.001; p=0.007; respectively). The POMA scores including both gait and balance components improved after the first and second procedures (p<0.05; for each). After the third procedure, the increase in POMA-balance score was statistically significant (p<0.05). After the first procedure, the FAB scores and NHPT scores were significantly improved (p<0.02). The median follow-up duration of the patients was 34.5 months. The deterioration of gait disturbance may be improved, and cognitive decline may be stabilized, at least postponed, by applying recurrent CSF removal in those unshunted patients with iNPH.


Multiple analyses of G-protein coupled receptor (GPCR) expression in the development of gefitinib-resistance in transforming non-small-cell lung cancer.
There is increasing evidence that functional crosstalk between GPCRs and EGFR contributes to the progression of colon, lung, breast, ovarian, prostate and head and neck tumors. In this study, we performed multiple analyses of GPCR expression in a gefitinib-resistant non-small cell lung cancer (NSCLC) cell line, H1975, which harbors an L858R/T790M mutation. To determine the expression profile of mRNAs encoding 384 GPCRs in normal human lung fibroblast (NHLF) and H1975 cells, a GPCR-specific microarray analysis was performed. A heat-map of the microarray revealed considerable differences in the expression of GPCRs between NHLF and H1975 cells. From the GPCR expression list, we selected some GPCR agonists/antagonist to investigate whether the respective ligands could affect the growth of H1975 cells. Among them, treatment with either a selective antagonist of adenosine A2a receptors, which were highly expressed in H1975 cell and another gefitinib-resistant NSCLC cells, HCC827GR cells or "small interfering RNA" (siRNA) targeting adenosine A2a receptors produced a significant decrease in cell viability of both H1975 and HCC827GR cells. Among up-regulated GPCRs in H1975 cells, Gs-, Gi- and Gq-coupled GPCRs were expressed almost equally. Among down-regulated GPCRs, Gi-coupled GPCRs were dominantly expressed in H1975 cells. The present results suggest that multilayered crosstalk between GPCRs and EGFR may play an important role in orchestrating downstream signaling molecules that are implicated in the development of gefitinib-resistant NSCLC.


The functional genomic distribution of protein divergence in two animal phyla: coevolution, genomic conflict, and constraint.
We compare the functional spectrum of protein evolution in two separate animal lineages with respect to two hypotheses: (1) rates of divergence are distributed similarly among functional classes within both lineages, indicating that selective pressure on the proteome is largely independent of organismic-level biological requirements; and (2) rates of divergence are distributed differently among functional classes within each lineage, indicating species-specific selective regimes impact genome-wide substitutional patterns. Integrating comparative genome sequence with data from tissue-specific expressed-sequence-tag (EST) libraries and detailed database annotations, we find a functional genomic signature of rapid evolution and selective constraint shared between mammalian and nematode lineages despite their extensive morphological and ecological differences and distant common ancestry. In both phyla, we find evidence of accelerated evolution among components of molecular systems involved in coevolutionary change. In mammals, lineage-specific fast evolving genes include those involved in reproduction, immunity, and possibly, maternal-fetal conflict. Likelihood ratio tests provide evidence for positive selection in these rapidly evolving functional categories in mammals. In contrast, slowly evolving genes, in terms of amino acid or insertion/deletion (indel) change, in both phyla are involved in core molecular processes such as transcription, translation, and protein transport. Thus, strong purifying selection appears to act on the same core cellular processes in both mammalian and nematode lineages, whereas positive and/or relaxed selection acts on different biological processes in each lineage.


Is concern about young people's anti-social behaviour associated with poor health? Cross-sectional evidence from residents of deprived urban neighbourhoods.
Young people in disadvantaged neighbourhoods are often the focus of concerns about anti-social behaviour (ASB). There is inconsistent evidence to support the hypothesis that perceptions of ASB (PASB) are associated with poor health. We ask whether perceptions of young people's ASB are associated with poor health; and whether health, demographic and (psycho)social characteristics can help explain why PASB varies within disadvantaged neighbourhoods (Glasgow, UK). Regression analysis of survey data exploring associations between perceiving teenagers hanging around to be a serious neighbourhood problem and SF-12v2 mental and physical health scores (higher = better), including adjustment for demographic characteristics. Further analysis explored associations with self-reported measures of health service use, psychosocial characteristics of homes and neighbourhoods and social contacts. 6008 adults participated (50% response) and 22% (n = 1,332) said teenagers were a serious neighbourhood problem (the most frequently reported local problem). Demographic characteristics associated with perceiving serious teenager problems included regular health service use, age (inverse relationship), financial problems and living with children. Lower SF-12v2 physical health scores were associated with perceiving teenager problems after adjustment for demographic variables (OR 0.98; 95%CI 0.97,0.99; p = < 0.001), whilst adjusted findings for mental health scores were less conclusive (OR 0.99; 95%CI 0.98,1.00; p = 0.103). Further analysis suggested that perceiving teenager problems was more strongly associated with a number of self-reported psychosocial factors: e.g. lacking social support, < weekly family contacts, poor neighbourhood safety, low trust in neighbours, neighbourhood perceived to be a barrier to self-esteem, and neighbourhood decline. Given the evidence we found of weak and small associations between PASB and health, we caution against assuming that tackling concern about teenagers' ASB will lead to substantial public health gains in disadvantaged areas. Although the findings do not present a compelling case for making PASB a public health priority, it is still important to address concerns about young people's ASB. Reasons for doing so may include improving social cohesion, reducing fear and isolation, and improving the general quality of people's lives - particularly in neighbourhoods burdened by multiple disadvantages. Future research should evaluate interventions that attempt to reduce PASB in disadvantaged areas. Findings from this study could help inform the targeting of such interventions.


Exploring short intramolecular interactions in alkylaromatic substrates.
From proteins and peptides to semiconducting polymers, aliphatic chains on aromatic groups are recurring motifs in macromolecules from very diverse application fields. Fields in which molecular folding and packing determine the macroscopic physical properties that make such advanced materials appealing in the first place. Within each macromolecule, the intrinsic structure of each unit defines how it interacts with its neighbours, ultimately opening up or denying certain backbone conformations. This eventually also determines how macromolecules interact with each other. This account deals specifically with the conformational problem of many common alkylaromatic units, examining the features of an intramolecular interaction involving a side chain with as few as three methylene groups. A set of 23 model compounds featuring an intramolecular interaction between an aliphatic X-H (X = C, N, O, and S) bond and an aromatic ring was considered. Quantitative computational analysis was made possible, thanks to complete basis set extrapolated CCSD(T) calculations and NCI topological analysis, the latter of which revealed an elaborate network of dispersive and steric interactions leading to somewhat unintuitive and unexpected results, such as the higher energetic stability of certain twisted conformational isomers over those with extended side chains. Vicinal covalent effects from polarizing groups and various heteroatoms, along with the occurrence of non-dispersive phenomena, were also investigated. The conclusions drawn from the investigation include a comprehensive set of guidelines intended to aid in the prediction of the most stable conformation for this class of building blocks. Our findings affect a variety of different research fields, including the tailoring of functional materials for organic electronics and photovoltaics, with insights into a rational treatment of conformational disorder, and the study of protein- and peptide-folding preferences, putting an emphasis on peculiar interactions between the backbone and aromatic residues.


PET-CT as an effective imaging modality in the staging and follow-up of post-transplant lymphoproliferative disorder following solid organ transplantation.
To establish the role of positron-emission tomography (PET)-computed tomography (CT) in post-transplant lymphoproliferative disorder (PTLD) patients, compared to conventional imaging (ultrasonography/CT/magnetic resonance imaging) in relation to its accuracy, sensitivity and specificity. 30 patients (26 males and 4 females), with a median age of 49.5 (range 18-74) years, were retrospectively evaluated. In 29 cases, the diagnosis was confirmed by histopathology. Malignant lymphoma was detected in 20 cases, polymorphic lymphoproliferative disorder in six cases, multiple myeloma in two cases and Hodgkin's disease in one case. A total of 49 PET-CTs (13 studies for staging at diagnosis and 36 studies at follow-up as assessment post-therapy) were compared to conventional imaging. Imaging results in accordance with disease status were assessed at a median follow-up of 17.8 (range 1.5-42.2) months post-PET-CT. In 41 of 49 examinations performed for staging and on follow-up, PET-CT and conventional imaging findings were concordant. Compared to conventional imaging, PET-CT showed comparable sensitivity (75 percent vs. 83 percent), similar specificity (100 percent in both modalities) and comparable accuracy (77 percent vs. 85 percent) during staging at diagnosis. PET-CT was found to be superior to conventional imaging modalities at follow-up, with greater sensitivity (100 percent vs. 81 percent), specificity (80 percent vs. 100 percent) and accuracy (97 percent vs. 83 percent). PET-CT is an accurate diagnostic tool for staging and for the follow-up of PTLD patients. It represents a good alternative imaging method to avoid contrast-related nephrotoxicity in patients who often develop impaired renal function secondary to chronic immunosuppressive therapy. However, further studies are recommended before considering PET-CT as a routine diagnostic tool in PTLD.


Young adult predictors of alcohol dependence to age 53: a 44-year prospective cohort study of Danish men.
To examine if (1) there is a positive association between drinking volume in young men and life-time risk of alcohol dependence (AD) and (2) there are other associations between young adulthood factors and life-time risk of AD. Prospective cohort study of sons of fathers with alcohol use disorder (AUD) and matched low-risk controls without paternal AUD. Setting and participants A total of 204 men, who were assessed at baseline in 1979 at age 19-20 years, were followed through record linkage with Danish registers and consecutive psychiatric interviews at the ages of 33, 43 and 53 years. AD diagnoses were interview-based according to the Diagnostic and Statistical Manual of Mental Disorders, 3rd edition, or made by treating clinicians according to the International Classification of Diseases (ICD) revision 8 (ICD-8) until 1993 and revision 10 (ICD-10) from 1994.We estimated odds ratios (ORs) with 95% confidence intervals (CI) for the development of AD after adjustment for confounders including smoking, social status and paternal AUD. The following variables from the examination at age 19-20 independently predicted life-time AD: alcohol consumption > 21 beverages/week versus 0-21 [odds ratio (OR) = 2.46, 95% confidence interval (CI) = 1.22-4.97], police contact (OR = 2.60, 95% CI = 1.28-5.28) and institutionalization related to the individual (OR = 2.90, 95% CI = 1.39-6.02). Compared with < 1 beverages/week, the risk for AD did not increase significantly for drinking volume categories: 1-7, 8-14 or 15-21 beverages/week. Independently of other risk factors in young adulthood, young Danish men's risk for life-time alcohol dependence appears to be predicted by a drinking volume at age 19-20 years exceeding 21 beverages per week.


Somatostatin and insulin mediate glucose-inhibited glucagon secretion in the pancreatic α-cell by lowering cAMP.
The dysregulation of glucose-inhibited glucagon secretion from the pancreatic islet α-cell is a critical component of diabetes pathology and metabolic disease. We show a previously uncharacterized [Ca(2+)]i-independent mechanism of glucagon suppression in human and murine pancreatic islets whereby cAMP and PKA signaling are decreased. This decrease is driven by the combination of somatostatin, which inhibits adenylyl cyclase production of cAMP via the Gαi subunit of the SSTR2, and insulin, which acts via its receptor to activate phosphodiesterase 3B and degrade cytosolic cAMP. Our data indicate that both somatostatin and insulin signaling are required to suppress cAMP/PKA and glucagon secretion from both human and murine α-cells, and the combination of these two signaling mechanisms is sufficient to reduce glucagon secretion from isolated α-cells as well as islets. Thus, we conclude that somatostatin and insulin together are critical paracrine mediators of glucose-inhibited glucagon secretion and function by lowering cAMP/PKA signaling with increasing glucose.


Comparative Sensitivity of Transthoracic and Transesophageal Echocardiography in Diagnosis of Infective Endocarditis Among Veterans With Staphylococcus aureus Bacteremia.
Echocardiography is fundamental for diagnosing infective endocarditis (IE) in patients with Staphylococcus aureus bacteremia (SAB), but whether all such patients require transesophageal echocardiography (TEE) is controversial. We identified SAB cases between February 2008 and April 2012. We compared sensitivity and specificity of transthoracic echocardiography (TTE) and TEE for evidence of IE, and we determined impacts of IE risk factors and TTE image quality on comparative sensitivities of TTE and TEE and their impact on clinical decision making. Of 215 evaluable SAB cases, 193 (90%) had TTE and 130 (60%) had TEE. In 119 cases with both tests, IE was diagnosed in 29 (24%), for whom endocardial involvement was evident in 25 (86%) by TEE, vs only 6 (21%) by TTE (P < .001). Transesophageal echocardiography was more sensitive than TTE regardless of risk factors. Even among the 66 cases with adequate or better quality TTE images, sensitivity was only 4 of 17 (24%) for TTE, vs 16 of 17 (94%) for TEE (P < .001). Among 130 patients with TEE, the TEE results, alone or with TTE results, influenced treatment duration in 56 (43%) cases and led to valve surgery in at least 4 (6%). It is notable that, despite vigorous efforts to obtain both tests routinely, TEE was not done in 86 cases (40%) for various reasons, including pathophysiological contraindications (14%), patient refusal or other patient-related factors (16%), and provider declination or system issues (10%). Patients with SAB should undergo TEE when possible to detect evidence for IE, especially if the results might affect management.


Evaluation of tumor necrosis factor-α (TNF-α) levels in plasma and their correlation with periodontal status in obese and non-obese subjects.
Obesity is emerging as a significant health problem worldwide and is a risk factor for various systemic diseases. Periodontal disease is a multifactorial inflammatory disease. Recent evidence points to a link between obesity and periodontal disease, and a role for tumor necrosis factor-α (TNF-α) has been suggested. FORTY NONDIABETIC SUBJECTS WERE DIVIDED INTO TWO GROUPS: group A (non-obese) included subjects with body mass index (BMI) of 18.5 to 27 kg/m2, and group B (obese) included subjects with BMI>27 kg/m2. The BMI, TNF-α levels in plasma and Periodontal Disease Index (PDI) scores were assessed, compared and correlated. Significantly higher PDI scores and TNF-α levels were found in the obese group as compared to those in the non-obese group. Also, a significant and positive correlation was seen between BMI and TNF-α, TNF-α and PDI as well as BMI and PDI. Increase in the levels of tumor necrosis factor-a in plasma and an increase in the severity of periodontal disease may be seen in subjects with a higher body mass index (BMI). This indicates that obesity may be detrimental to the periodontal health of individuals.


Do new guidelines and technology make the routine pelvic examination obsolete?
Routine pelvic examinations are the core of the periodic gynecological examination and widely tolerated as a necessary part of health maintenance. Is this examination beneficial for asymptomatic women? Justifications for the pelvic examination include screening for Chlamydia (or gonorrhea) infection, evaluation before initiation of hormonal contraception, screening for cervical cancer, and early detection of ovarian cancer. Current nucleic acid amplification tests for Chlamydia and gonorrhea permit the use of urine and self-administered vaginal swabs, which most women prefer over a pelvic examination. Pelvic examination findings do not affect the decision to prescribe or withhold systemic hormonal contraception; a pelvic examination is not needed to initiate these contraceptives. Recent American College of Obstetricians and Gynecologists (ACOG) guidelines recommend less frequent cervical screening, thus decreasing the frequency of a speculum examination for cervical screening. Bimanual examinations for palpation of the uterus and ovaries are also routinely performed in the United States. Clinical trial data, however, show these examinations do not lead to earlier detection of ovarian cancer. No evidence identifies benefits of a pelvic examination in the early diagnosis of other conditions in the asymptomatic woman. Speculum and bimanual examinations are uncomfortable, disliked by many women, and use scarce time during a well woman visit. Eliminating the speculum examination from most visits and the bimanual examination from all visits of asymptomatic women will free resources to provide services of proven benefit. Overuse of the pelvic examination contributes to high healthcare costs without any compensatory health benefit.


Backgrounds and characteristics of arsonists.
The aim of this study was to gain more insight in the backgrounds and characteristics of arsonists. For this, the psychiatric, psychological, personal, and criminal backgrounds of all arsonists (n=25), sentenced to forced treatment in the maximum security forensic hospital "De Kijvelanden", were compared to the characteristics of a control group of patients (n=50), incarcerated at the same institution for other severe crimes. Apart from DSM-IV Axis I and Axis II disorders, family backgrounds, level of education, treatment history, intelligence (WAIS scores), and PCL-R scores were included in the comparisons. Furthermore, the apparent motives for the arson offences were explored. It was found that arsonists had more often received psychiatric treatment, prior to committing their index offence, and had a history of severe alcohol abuse more often in comparison to the controls. The arsonists turned out to be less likely to suffer from a major psychotic disorder. Both groups did not differ significantly on the other variables, among which the PCL-R total scores and factor scores. Exploratory analyses however, did suggest that arsonists may differentiate from non-arsonists on three items of the PCL-R, namely impulsivity (higher scores), superficial charm (lower scores), and juvenile delinquency (lower scores). Although the number of arsonists with a major psychotic disorder was relatively low (28%), delusional thinking of some form was judged to play a role in causing arson crimes in about half of the cases (52%).


Prevalence of illicit drugs detected in the urine of women of childbearing age in Alabama public health clinics.
Each year, it is estimated that from 350,000 to 739,000 U.S. infants are exposed in utero to one or more illicit drugs. To estimate the prevalence of and risk factors for illicit drug use by women of childbearing age in Alabama, during 2 months in 1991 the authors collected patient-reported histories, clinical histories, and urine specimens from 6,195 women statewide attending public health maternity clinics, family planning clinics, and a high-risk referral obstetrical clinic. Blind drug screening of urine specimens for marijuana, cocaine, opiates, barbiturates, and amphetamines was performed with the use of a fluorescent polarization immunoassay. The overall prevalence of positive results for drugs tested was 10.1 percent, including 8.4 percent of the 3,554 pregnant and 12.3 percent of the 2,571 nonpregnant women screened. The drugs most frequently detected were marijuana and cocaine. Characteristics of the subjects associated with a higher prevalence of positive results for any drug tested or for marijuana included white race, older age, being divorced, non student occupation, having 12 or less years of education, attending a clinic located in a suburban county, self-reported substance use, increased risk for human immunodeficiency virus infection, and reproductive history. Characteristics of women with positive screening for cocaine results were similar to those who tested positive for any drug, except that the prevalence of cocaine was higher among black women and those attending urban county clinics and did not vary by years of education. Patient-reported histories of drug use were insensitive in identifying women who had positive drug screening results (sensitivity, 6.3 percent; specificity, 98.2 percent). Thus, in this study,the use of illicit drugs among women of childbearing age attending public clinics in Alabama was common and emphasizes the need for targeted drug education and interventions to reduce the impact of drug use on this high-risk population.


Tuning the affinity of catechols and salicylic acids towards Al(iii): characterization of Al-chelator interactions.
Due to aluminum's controversial role in neurotoxicity, the goal of chelation therapy, the removal of the toxic metal ion or attenuation of its toxicity by transforming it into less toxic compounds, has attracted considerable interest in the past years. In the present paper we present, validate and apply a state-of-the-art theoretical protocol suitable for the characterization of the interactions between a chelating agent and Al(iii). In particular, we employ a cluster-continuum approach based on Density Functional Theory calculations to evaluate the binding affinity of aluminum for a set of two important families of aromatic chelators: salicylic acids and catechols. Our protocol shows very good qualitative agreement between the computed binding affinities and available experimental stability constants (log β) values for 1 : 1, 1 : 2 and 1 : 3 complexes. Then, we have investigated the nature of the Al-O bond in an enlarged dataset of 27 complexes of 1 : 1 stoichiometry, by means of the QTAIM and Energy Decomposition Analysis (EDA). Quite interestingly, we have found that although the Al-O interaction is mainly electrostatic, there is a small but significant degree of covalency that explains the modulation of binding affinities in both families of compounds by the addition of electron donating (CH3, OCH3) or withdrawing (NO2, CF3) substituents. The role of aromaticity and the mechanisms of action of the different functional groups were also evaluated. Finally, we have analyzed the competition between Al(iii) and proton toward the binding of these chelators, giving a rationalization of the different trends found experimentally between log β and the amount of free aluminum in solution in the presence of a given ligand (p[Al]). In summary, we propose a validated and comprehensive computational protocol that can provide a valuable help toward the design and tuning of new efficient aluminum chelators.


Influence of calcareous soil on Cry3Bb1 expression and efficacy in the field.
Greater than expected injury by western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte) to Cry3Bb1 expressing maize hybrids (Zea mays L.) has been reported in southwestern Nebraska. Affected areas of some fields are often associated with high pH calcareous soils where maize growth is poor and iron chlorosis is common. As part of a comprehensive study to understand potential causes of unexpected injury, experiments were conducted during 2013 and 2014 to ascertain whether the calcareous soil conditions and associated poor maize growth negatively affect the expression of Cry3Bb1. Quantitative determination of Cry3Bb1 protein expression levels in root tissues was carried out on plants at V5-V6 growth stage using the enzyme-linked immunosorbent assay. Cry3Bb1 and non-Bt near isoline maize hybrids were artificially infested with Cry3Bb1-susceptible WCR eggs to measure survival and efficacy of Cry3Bb1 maize in calcareous and non-calcareous soils. Results showed that there was not a significant difference in expression of Cry3Bb1 protein between plants from calcareous and non-calcareous soils (18.9-21.2 µg/g fresh weight). Western corn rootworm survival was about sevenfold greater from the non-Bt isoline than Cry3Bb1 maize indicating that Cry3Bb1 performed as expected when infested with a Cry3Bb1 susceptible rootworm population. When survival from calcareous and non-calcareous soils was compared, no significant differences were observed in each soil. A significant positive correlation between soil pH and expression of Cry3Bb1 protein in roots was detected from samples collected in 2014 but not in 2013. No such correlation was found between soil pH and survival of WCR. Results suggest that Cry3Bb1 expression levels were sufficient to provide adequate root protection against WCR regardless of soil environment, indicating that lowered Cry3Bb1 expression is not a contributing factor to the greater than expected WCR injury observed in some southwestern Nebraska maize fields.


Cat Ownership Perception and Caretaking Explored in an Internet Survey of People Associated with Cats.
People who feed cats that they do not perceive they own (sometimes called semi-owners) are thought to make a considerable contribution to unwanted cat numbers because the cats they support are generally not sterilized. Understanding people's perception of cat ownership and the psychology underlying cat semi-ownership could inform approaches to mitigate the negative effects of cat semi-ownership. The primary aims of this study were to investigate cat ownership perception and to examine its association with human-cat interactions and caretaking behaviours. A secondary aim was to evaluate a definition of cat semi-ownership (including an association time of ≥1 month and frequent feeding), revised from a previous definition proposed in the literature to distinguish cat semi-ownership from casual interactions with unowned cats. Cat owners and semi-owners displayed similar types of interactions and caretaking behaviours. Nevertheless, caretaking behaviours were more commonly displayed towards owned cats than semi-owned cats, and semi-owned cats were more likely to have produced kittens (p<0.01). All interactions and caretaking behaviours were more likely to be displayed towards cats in semi-ownership relationships compared to casual interaction relationships. Determinants of cat ownership perception were identified (p<0.05) and included association time, attachment, perceived cat friendliness and health, and feelings about unowned cats, including the acceptability of feeding unowned cats. Encouraging semi-owners to have the cats they care for sterilized may assist in reducing the number of unwanted kittens and could be a valuable alternative to trying to prevent semi-ownership entirely. Highly accessible semi-owner "gatekeepers" could help to deliver education messages and facilitate the provision of cat sterilization services to semi-owners. This research enabled semi-ownership to be distinguished from casual interaction relationships and can assist welfare and government agencies to identify cat semi-owners in order to develop strategies to address this source of unwanted cats.


Study Design and qPCR Data Analysis Guidelines for Reliable Circulating miRNA Biomarker Experiments: A Review.
In the past decade, the search for circulating microRNA (miRNA) biomarkers has yielded numerous associations between miRNAs and different types of disease. However, many of these relations could not be replicated in subsequent studies under similar experimental conditions. Although this lack of replicability may be explained by the variation in experimental design and analysis methods, guidelines on the most appropriate design and analysis methods to study circulating miRNAs are scarce. miRNA biomarker experiments generally consist of a discovery phase and a validation phase. In the discovery phase, typically hundreds of miRNAs are measured in parallel to identify candidate biomarkers. Because of the costs of such high-throughput experiments, the number of individuals included in those studies is often too small, which can easily lead to false positives and false negatives. In the validation phase, a small number of identified biomarker candidates are measured in a large cohort of cases and controls, generally by quantitative PCR (qPCR). Although qPCR is a sensitive method to measure miRNAs in the circulation, experimental design and qPCR data analysis remain challenging. Omitting some crucial steps in the design and analysis of the qPCR experiment or performing them incorrectly can cause serious biases, ultimately leading to false conclusions. In this review, we aim to expose and discuss the most common sources of interstudy variation in miRNA research from a methodological point of view and to provide guidelines on how to perform these steps correctly to increase replicability of studies on circulating miRNAs.